CN111812054A - System and method for detecting propofol online derivatization of optical fiber sensing microfluidic chip - Google Patents
System and method for detecting propofol online derivatization of optical fiber sensing microfluidic chip Download PDFInfo
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- CN111812054A CN111812054A CN202010724659.7A CN202010724659A CN111812054A CN 111812054 A CN111812054 A CN 111812054A CN 202010724659 A CN202010724659 A CN 202010724659A CN 111812054 A CN111812054 A CN 111812054A
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- 239000013307 optical fiber Substances 0.000 title claims abstract description 61
- 238000001212 derivatisation Methods 0.000 title claims abstract description 60
- OLBCVFGFOZPWHH-UHFFFAOYSA-N propofol Chemical compound CC(C)C1=CC=CC(C(C)C)=C1O OLBCVFGFOZPWHH-UHFFFAOYSA-N 0.000 title claims abstract description 46
- 229960004134 propofol Drugs 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 21
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- WGTYBPLFGIVFAS-UHFFFAOYSA-M tetramethylammonium hydroxide Chemical compound [OH-].C[N+](C)(C)C WGTYBPLFGIVFAS-UHFFFAOYSA-M 0.000 claims description 6
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/50273—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- B—PERFORMING OPERATIONS; TRANSPORTING
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Abstract
The invention discloses the technical field of rapid detection of medicaments, in particular to an on-line derivatization detection system and a method of propofol of an optical fiber sensing micro-fluidic chip, wherein the system comprises an injection pump, a micro-fluidic mixed chip and a Z-shaped flow cell, the micro-fluidic mixed chip is adopted for carrying out on-line derivatization on a sample and a derivatization reagent, and the flow velocity and the flow quantity of the reagent can be accurately controlled, so that the detection can be finished by using trace samples and reagents; the optical fiber sensing technology is adopted for on-line detection, optical signals are transmitted through optical fibers, the structure of the instrument is simplified, the volume of the instrument is reduced, the whole analysis system is miniaturized and portable, and the requirements of on-site rapid detection are met; the on-line derivatization technology is adopted to carry out rapid detection on the biological sample, the detection sensitivity and accuracy are improved through derivatization, and the problem that the spectrum detection is easily interfered by other impurities in the sample is solved.
Description
Technical Field
The invention relates to the technical field of rapid detection of medicines, in particular to a system and a method for detecting propofol online derivatization by using an optical fiber sensing microfluidic chip.
Background
Immediate testing of narcotics has long been a clinical need, with overdose resulting in circulation and respiratory depression, and under-dose resulting in sensory pain to the patient. At present, the control and adjustment of the drug dosage mainly depend on clinical experience and the monitoring of physical signs of patients, the drug metabolism condition has larger individual difference, the control and adjustment of the drug dosage are important for the reasonability, safety and effectiveness of the drug administration, the key point of the control and adjustment depends on the feedback of the in-vivo behavior of the individual drug in time, and the in-vivo drug concentration is the most important auxiliary information. Due to the limitation of characteristics such as a rigid structure of an analysis instrument, the existing in-vivo drug concentration analysis method is offline sampling analysis and detection, the process is complex and time-consuming, the analysis result is not easy to obtain in time, the problem of in-vivo drug information feedback lag exists, and the existing anesthesia depth control lacks of better objective evaluation.
A new technology and a new method for the instant detection of in vivo drug components are hot spots of the current domestic and foreign researches, a feasible instant detection method for anesthetic drugs is not available at present, and an optical fiber sensing microfluidic chip propofol online derivatization detection system and method are provided.
Disclosure of Invention
The invention aims to provide a modern intravenous anesthetic propofol detection system and method which are most widely applied at home and abroad, and by combining a microfluidic chip technology, an optical fiber chemical sensing online process monitoring technology and an online derivatization technology, a miniaturized, online and field measurement analysis system is constructed, a new method for rapidly detecting the concentration of an anesthetic in real time online is established, the in-vivo behavior of the detected drug is evaluated, the safety and the effectiveness of clinical anesthetic administration are further improved, and a new technology and a new idea for researching a more rapid, real-time and online high-sensitivity drug analysis are developed.
In order to achieve the purpose, the invention provides the following technical scheme: the on-line derivatization detection system for propofol of the optical fiber sensing micro-fluidic chip comprises an injection pump, wherein the outlet end of the injection pump is connected with the inlet end of a micro-fluidic mixing chip, the outlet end of the micro-fluidic mixing chip is connected with the liquid inlet of a Z-shaped flow cell, the liquid outlet of the Z-shaped flow cell is connected with a waste liquid container through a connecting pipe, a light path inlet and a light path outlet are respectively arranged on the Z-shaped flow cell, the light path inlet is connected with a light source through an input optical fiber, the light path outlet is connected with an optical fiber spectrometer through an output optical fiber, the optical fiber spectrometer is electrically connected with a computer, and spectrum acquisition and processing software.
Preferably, be provided with the Z shape pipeline in the Z type flow-through cell, two port departments of Z shape pipeline do respectively the inlet and the liquid outlet of Z type flow-through cell, the department of buckling of Z shape pipeline does respectively the light path entry with the light path export.
Preferably, the working process of the on-line propofol derivatization detection system of the optical fiber sensing microfluidic chip is as follows: a derivatization reagent and a propofol sample reagent enter the microfluidic mixing chip through the injection pump to carry out online derivatization reaction, a derivatization product is generated and then is subjected to ultraviolet-visible spectrum real-time detection through the Z-shaped flow cell, an optical fiber sensor is adopted to transmit an optical signal, light of the light source is guided into the Z-shaped flow cell through the input optical fiber, the light is guided into the optical fiber spectrometer through the output optical fiber to carry out spectrum detection, the optical fiber spectrometer transmits the signal into a computer, and spectrum acquisition and processing software obtains a real-time dynamic change ultraviolet-visible spectrum diagram of the derivatization reaction.
Preferably, the injection pump is an independently controllable multi-channel injection pump, a plurality of channels can be provided for accurately controlling the flow speed and the flow rate of a detected sample and a derivatization reagent, the multi-channel injection pump can adopt a TS-1B injection pump, the TS-1B injection pump is a controller of a four-channel multifunctional injection pump, different working parameters can be set for the four channels respectively, the multi-channel multi-functional injection pump can be provided with standard injectors with various specifications of 5 mu L-60 mL, and the requirements of different experiments can be met by accurate stroke control and ultra-wide range linear speed (7.9 mu m/min-79.4 mm/min).
Preferably, the chip of the microfluidic mixing chip is formed by bonding an upper substrate and a lower substrate, wherein the upper substrate is made of polydimethylsiloxane, the lower substrate is made of glass, the depth and width of a microchannel of the microfluidic mixing chip are both 50-500 μm, the mixing chip is provided with three inlets and one outlet, a mixing pipeline is designed by adopting an S-shaped pipeline, and the mixing chip is used for fully mixing sample test solution and a derivatization reagent in the microchannel to generate a sample derivatization product, so that the online derivatization treatment of the sample is realized.
Preferably, the light source is a deuterium-tungsten halogen combined light source which can provide stable and continuous spectral output in an ultraviolet-visible spectrum region, and the deuterium-tungsten halogen combined light source can adopt a DH-2000-BAL deuterium-tungsten halogen combined light source which can generate stable and continuous spectral output at 215nm-2000 nm.
Preferably, the optical fiber spectrometer adopts a back-illuminated CCD detector to measure the absorbance change of the sample derivatization product in real time, and the back-illuminated CCD detector can adopt a Maya 2000PRO optical fiber spectrometer to detect the absorbance within the wavelength range of 200nm-900 nm.
Preferably, the input optical fiber (8) and the output optical fiber (10) are both transmitted by adopting ultraviolet-visible optical fibers, the input optical fiber and the output optical fiber are both QP400-2-UV-VIS optical fibers, the length of the optical fibers is 2m, the diameter of the optical fibers is 400 mu m, and the optical fibers are suitable for detecting ultraviolet light and visible light.
Preferably, the spectrum acquisition processing software acquires and records the spectrum signal of the sample derivative product generated on line on the microfluidic chip in real time to form a visual real-time dynamic change signal curve, and the spectrum acquisition processing software can adopt a spectra suite spectrum workstation to acquire the real-time spectrum of the sample measurement.
The method for detecting propofol online derivatization by using the optical fiber sensing microfluidic chip comprises the following steps: a Gibbs 'derivatization method is adopted, a Gibbs' reaction derivatization method is adopted for propofol detection, the structural modification of a derivatization reagent enables the detection wavelength of a derivatization product to be red-shifted to a visible light region, the interference of endogenous substances and exogenous substances in blood is eliminated, and the sensitivity and the selectivity of propofol ultraviolet-visible spectrum detection are improved
Gibbs' reaction principle: the Gibbs' reagent generates quinonimine under the alkaline condition provided by tetramethyl ammonium hydroxide solution, and the quinonimine and active hydrogen at the para-position of the phenolic hydroxyl of the propofol synthesize a blue compound. The reaction can be completed instantly at normal temperature, and is suitable for the requirements of on-line rapid derivatization and detection of propofol.
The reaction equation is as follows:
the Gibbs' reagent concentration is 0.15mg/mL, the concentration of the tetramethylammonium hydroxide solution is 0.05%, the sample introduction flow rates of the reagent and the sample are both 75 mu L/min, a stable derivatization product is obtained after 3min of derivatization reaction, the detection is carried out at 615nm, and the linear range of the propofol biological sample detection is 1.0-18.0 mu g/mL.
Compared with the prior art, the invention has the beneficial effects that: the system and the method for detecting propofol online derivatization by using the optical fiber sensing microfluidic chip have the following advantages:
1. the micro-fluidic mixing chip is adopted to carry out on-line derivatization on the sample and the derivatization reagent, and the flow velocity and the flow of the reagent can be accurately controlled, so that the detection can be finished by using trace samples and reagents.
2. The optical fiber sensing technology is adopted for on-line detection, optical signals are transmitted through the optical fibers, the structure of the instrument is simplified, the volume of the instrument is reduced, the whole analysis system is miniaturized and portable, and the on-site rapid detection requirement is met.
3. The on-line derivatization technology is adopted to carry out rapid detection on the biological sample, the detection sensitivity and accuracy are improved through derivatization, and the problem that the spectrum detection is easily interfered by other impurities in the sample is solved.
4. The optical fiber sensing micro-fluidic chip on-line derivatization detection system established by the invention can be used for the ultraviolet-visible spectrum rapid detection of various substances such as food, medicines, clinical biological samples, toxic and harmful components in the environment and the like, and has wide application prospect.
Drawings
FIG. 1 is a standard curve for propofol derivatization detection in an embodiment of the present invention;
FIG. 2 is a graph of UV-Vis spectra of propofol control and biological samples after derivatization in an embodiment of the invention, where 1 is a blank plasma sample, 2 is plasma to which propofol standard solution is added, and 3 is the propofol control;
fig. 3 is a schematic structural diagram of an optical fiber sensing microfluidic chip propofol online derivatization detection system in the embodiment of the invention.
In the figure: the system comprises a 1-injection pump, a 2-microfluidic mixing chip, a 3-Z type flow cell, a 4-connecting pipe, a 5-waste liquid container, a 6-light path inlet, a 7-light path outlet, an 8-input optical fiber, a 9-light source, a 10-output optical fiber, an 11-optical fiber spectrometer and a 12-computer.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example (b): referring to figures 1-3 of the drawings,
step 1: pretreatment of propofol in a biological sample:
precisely sucking 0.5mL of propofol containing plasma sample, diluting the sample with 4% phosphoric acid with the same volume, respectively activating the HLB solid-phase extraction column with 1mL of methanol, balancing 1mL of water, adding 1mL of plasma sample acidified by phosphoric acid, adding the solid-phase extraction column, leaching with 1mL of water, and adding 1mL of 1% KHCO3And (3) leaching the mixture twice by acetonitrile-water (1:9), finally eluting the mixture for 2 times by 0.5mL of methanol, and merging and collecting eluent.
Step 2: examination of the linear range:
490 mu L of propofol blank plasma 5 parts are precisely measured, 10 mu L of propofol series concentration standard solution is respectively added to make the blood concentration 1.0, 3.0, 6.0, 9.0, 12.0, 15.0 and 18.0 mu g/mL. The method comprises the following steps of 1, preprocessing a sample, performing online derivatization and detection by using an optical fiber sensing microfluidic chip propofol online derivatization detection system, determining the absorbance of the sample, performing linear regression by using a blank plasma sample as a blank control, using the propofol drug-containing plasma concentration (mu g/mL) as an abscissa X and the absorbance as an ordinate Y, wherein a standard curve equation is Y ═ 0.0283X +0.0812, and r ═ 0.9960, and the linear relation is good in a concentration range of 1.0-18.0 mu g/mL.
Propofol plasma concentration and absorbance
And step 3: recovery and precision
490 mu L of propofol blank plasma is precisely measured, 10 mu L of propofol series concentration standard solution is respectively added to ensure that the blood concentration is 5.0, 10.0 and 15.0 mu g/mL, 3 parts of each concentration sample are prepared in parallel, and the recovery rate of the sample is determined after the sample is pretreated by adopting the treatment method in the step 1.
Recovery and precision measurements (n ═ 3)
And 4, step 4: simulated biological sample assay
Accurately measuring 490 mu L of propofol blank plasma 9 parts, respectively adding propofol injection emulsion, calculating according to the marked amount to enable the blood concentration to be 4.0, 8.0 and 12.0 mu g/mL, preparing 3 parts for each concentration in parallel, carrying out sample pretreatment by adopting the treatment method in the step 1, carrying out online derivatization and detection by using an optical fiber sensing microfluidic chip propofol online derivatization detection system, and calculating the propofol concentration according to a standard curve equation.
Simulation of biological sample measurement results (n ═ 3)
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. The on-line derivatization detection system for propofol of the optical fiber sensing microfluidic chip is characterized in that: including injection pump (1), the exit end of injection pump (1) is connected with the entrance point of micro-fluidic mixed chip (2), the exit end of micro-fluidic mixed chip (2) with the inlet of Z type flow cell (3) is connected, the liquid outlet of Z type flow cell (3) is connected with waste liquid container (5) through connecting pipe (4), be provided with light path entry (6) and light path export (7) on Z type flow cell (3) respectively, light path entry (6) are connected with light source (9) through input optical fiber (8), light path export (7) are connected with fiber optic spectrometer (11) through output optical fiber (10), fiber optic spectrometer (11) are connected with computer (12) electricity, install spectrum collection processing software in computer (12).
2. The system for detecting propofol online derivatization of an optical fiber sensing microfluidic chip according to claim 1, wherein the system comprises: be provided with Z shape pipeline in Z type flow-through cell (3), two port departments of Z shape pipeline do respectively the inlet and the liquid outlet of Z type flow-through cell (3), the department of buckling of Z shape pipeline does respectively light path entry (6) with light path export (7).
3. The system for detecting propofol online derivatization of an optical fiber sensing microfluidic chip according to claim 1 or 2, wherein the system comprises: the working process is as follows: a derivatization reagent and a propofol sample reagent enter the microfluidic mixing chip (2) through the injection pump (1) to carry out online derivatization reaction, ultraviolet-visible spectrum real-time detection is carried out through the Z-shaped flow cell (3) after a derivatization product is generated, an optical fiber sensor is adopted to transmit optical signals, light of the light source (9) is guided into the Z-shaped flow cell (3) through the input optical fiber (8), the light is guided into the optical fiber spectrometer (11) through the output optical fiber (10) to carry out spectrum detection, the optical fiber spectrometer (11) transmits the signals into the computer (12), and spectrum acquisition and processing software obtains a real-time dynamic change ultraviolet-visible spectrum diagram of the derivatization reaction.
4. The system for detecting propofol online derivatization of an optical fiber sensing microfluidic chip according to claim 3, wherein the system comprises: the injection pump (1) is a multi-channel injection pump which can be independently controlled.
5. The system for detecting propofol online derivatization of an optical fiber sensing microfluidic chip according to claim 3, wherein the system comprises: the chip of the microfluidic mixing chip (2) is formed by bonding an upper substrate and a lower substrate, wherein the upper substrate is made of polydimethylsiloxane, the lower substrate is made of glass, the depth and width of a microchannel of the microfluidic mixing chip (2) are 50-500 micrometers, the mixing chip is composed of three inlets and one outlet, and the mixing pipeline is an S-shaped pipeline.
6. The system for detecting propofol online derivatization of an optical fiber sensing microfluidic chip according to claim 3, wherein the system comprises: the light source (9) is a deuterium-tungsten halogen combined light source.
7. The system for detecting propofol online derivatization of an optical fiber sensing microfluidic chip according to claim 3, wherein the system comprises: the fiber spectrometer (11) adopts a back-illuminated CCD detector.
8. The system for detecting propofol online derivatization of an optical fiber sensing microfluidic chip according to claim 3, wherein the system comprises: the input optical fiber (8) and the output optical fiber (10) are both transmitted by adopting ultraviolet-visible optical fibers.
9. The system for detecting propofol online derivatization of an optical fiber sensing microfluidic chip according to claim 3, wherein the system comprises: the spectrum acquisition processing software acquires and records the spectrum signal of the sample derivatization product generated on line on the microfluidic chip in real time to form a visual real-time dynamic change signal curve.
10. The on-line derivatization detection method of propofol of the optical fiber sensing microfluidic chip is characterized by comprising the following steps: the method comprises the following steps: by adopting a Gibbs 'derivatization method, the concentration of a Gibbs' reagent is 0.15mg/mL, the concentration of a tetramethylammonium hydroxide solution is 0.05%, the sample introduction flow rates of the reagent and a sample are both 75 mu L/min, a stable derivatization product is obtained after derivatization reaction is carried out for 3min, detection is carried out at 615nm, and the linear range of propofol biological sample detection is 1.0-18.0 mu g/mL.
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